The SEM stubbs are always prepped dry.
You can read my materials and methods section soon.
Remember, these are puffballs. The spores are dry within the gleba when mature. If I used fresh, wet material, then the gleba would be immature, and therefore the spores immature as well.
Gasterocarp 10-35 mm tall x 12-40 mm broad; slightly obpyriform to compressed globose, often plicate at the base; rhizomorph up to 5 mm thick at the base where attached to the gasterocarp x 10 mm long, incrusted with vegetal debris and soil, rooting the base of the fruiting body to the soil, usually as a single thick strand with fine branches, or a tuft of compact mycelium made with tightly woven thin hyphal hairs, sometimes attaching several fruit bodies in a caespitose manner, eventually breaking off at the soil level to roll about the landscape; ostiole up to 4-5 mm broad at maturity, developing at the apex with the sloughing off of the exoperidium, irregularly torn to slit-like or circular, persistent, sometimes slightly upturned at the rim. Exoperidium cream white when young, becoming pale yellow white (3A2) to buff pink (5A3), often with pink grey tones near the apex (7A2), sometimes a greenish yellow (4D4), becoming buff beige (5C5) to brown gray (5C2) with maturity and sun exposure, often colored in a patchy or scale-like pattern; thin, seemingly glabrous at the apex, furfuraceous-verrucose to subflocculose-granulose at the base and in between folds, readily rubbing off, ornamentation obvious under a dissecting scope but not to the naked eye, wrinkled to folded near the base where plicate, sometimes with an areolate pattern when sun-exposed, cracking with desiccation to form a scale checkered-like pattern and slowly sloughing off in flakes, wearing away to expose the endoperidium with maturity. Endoperidium yellowish grey when young (3B2) to dull yellow
(3B3), light gray (5B2), turning grayish orange (5B3), to brownish yellow (5C4-7) to olive brown when dry (4E5) and often a darker red brown (8F6) at the base with maturity; thin, glabrous to smooth, parchment-like, persistent, dull or with a metallic sheen when very mature. Gleba white when young (5A1), turning grayish yellow (4B3), to brownish orange (5C4), then olive green (4F7), to dark chocolate brown (6F4) at maturity; firm and solid when young, cottony in texture with age, maturing from the center outward with the lower portions maturing last. Subgleba yellow cream buff (5A3) to brownish orange (5C3); cellular and composed of compact cells in smaller specimens, greatly reduced or absent in larger specimens as if stretched thin from growth, comprising only the lower portions of the gasterocarp when present. Diaphragm absent.
Basidiospores globose to subglobose; 3.5-6 X 3.5-5 µm [xmr = 4.5-4.9 X 4.4-4.5 µm, xmm = 4.8 ± 0.1 X 4.4 ± 0.1 µm, Q = 0.8-1.2, Qmr = 1.0-1.1, Qmm = 1.1 ± 0.0, n = 25, s = 8]; spores hyaline to golden brown in water mounts, turning bright green in KOH; seemingly smooth or slightly verrucose under light microscope, ornamentation can be seen under the light microscope using the lactic acid cotton blue reaction, with SEM spores mid to large verrucose warts, spaced apart and connected by thin rail-like bridges; central oil drop present; spores thick-walled; pedicel short and rudimentary, up to 0.8 μm long; free-floating sterigma not present in wet mounts; spores mostly of equal size under the light microscope. Eucapillitium Intermediate-type, threads 2-12 μm broad with walls 0.5-1.5 μm thick; hyaline to dark brown in water mounts, golden yellow to brown-green yellow in KOH, thin threads hyaline, thicker threads pigmented; elastic, breaking evenly where severed; glabrous to finely incrusted on thicker strands, sparse to frequent dichotomous branching, threads mostly straight, rarely undulate, sometimes subundulate in thinner threads, with knob-like projections; tips attenuate to fine rounded ends; Lycoperdon-type
sometimes found near the inner endoperidium wall in mature specimens. Small to medium-sized punctate pores abundant and up to 1 µm wide, pores more abundant on larger capillitial threads. Septa present and rare, pseudosepta present. Paracapillitium absent. Exoperidium textura angularis; composed of hyaline, irregular-shaped, tightly packed, inflated, thin-walled cells. Endoperidium textura intricata; composed of septate, thick-walled, tightly woven hyphal threads.
Habitat: A common puffball that is terrestrial and dispersed in open grassy terrain. Rarely found solitary, growing in numerous scattered clusters to gregarious or caespitose, and sometimes growing in ferry rings in wet years. Often fruiting in sandy, well-drained soil. Bovista aestivalis will fruit in mixed conifer forest, under Aspen (Populus tremuloides), in the Sonoran Desert Scrub, and with coastal oak species; suggesting a non-mycorrhizal ecology. Among the Geographic Subdivisions of California, this species can be found in the Cascade Range, the Modoc Plateau, in the Sierra Nevada, and in Central Western California.
But I wonder how much the SEM photo looks like it looks when fresh. How was the sample prepared? The SEM looks almost like a lunar landscape. Usually when I do microscopy everything is well hydrated, in these everything is completely dry. How much does the method you use for sample prep affect the results?
Created: 2009-06-10 20:26:41 CDT (-0500)
Last modified: 2014-02-26 11:44:44 CST (-0600)
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